Jin J P, Malik M L, Lin J J
Department of Biology, University of Iowa, Iowa City.
Hybridoma. 1990 Dec;9(6):597-608. doi: 10.1089/hyb.1990.9.597.
Three mouse IgG1 monoclonal antibodies (MAbs), named FA1, FA2, and FA3, against cardiac myosin heavy chain (MHC) with high specificity have been obtained. The immunogen used to generate these MAbs was the high-salt- and detergent-insoluble fraction of adult rat myocardial tissue. Western blots showed that these MAbs reacted with a 200 kD protein band, which comigrated with the heavy chain of purified rat cardiac myosin in SDS-PAGE. Immunofluorescence microscopy revealed that the antigen recognized by these MAbs was localized at the A-band of isolated myofibrils. The tissue-, species-, and isoform-specificities of these MAbs were examined by Western blots on various muscle samples. FA2 recognized fish, frog, chicken, rabbit, bovine, mouse and rat cardiac MHC, as well as rabbit skeletal and rat aorta smooth muscle MHC. This antibody reacted equally well with both alpha- and beta-isoforms of MHC. FA1 did not crossreact with any MHC tested so far but with rat cardiac MHC. It appeared to react only with alpha-isoform of MHC. FA3 recognized only rat, bovine and rabbit cardiac MHC with the specificity to bovine and rabbit atrial MHC. Elisa competition assay revealed that different epitopes on the antigen molecules were recognized by these three MAbs, although there was a partial overlap between the epitopes for FA1 and FA2. These anti-MHC MAbs will be most useful in investigating the expression of MHC during myocardial development.
已获得三种针对心肌肌球蛋白重链（MHC）的具有高特异性的小鼠IgG1单克隆抗体（MAb），分别命名为FA1、FA2和FA3。用于产生这些单克隆抗体的免疫原是成年大鼠心肌组织的高盐和去污剂不溶性部分。蛋白质免疫印迹显示，这些单克隆抗体与一条200 kD的蛋白带发生反应，该蛋白带在SDS-PAGE中与纯化的大鼠心肌肌球蛋白重链迁移位置相同。免疫荧光显微镜检查显示，这些单克隆抗体识别的抗原定位于分离的肌原纤维的A带。通过对各种肌肉样本进行蛋白质免疫印迹来检测这些单克隆抗体的组织、物种和同工型特异性。FA2识别鱼类、青蛙、鸡、兔、牛、小鼠和大鼠的心脏MHC，以及兔骨骼肌和大鼠主动脉平滑肌MHC。该抗体与MHC的α和β同工型反应同样良好。FA1到目前为止与任何测试的MHC都没有交叉反应，但与大鼠心脏MHC有交叉反应。它似乎只与MHC的α同工型反应。FA3只识别大鼠、牛和兔的心脏MHC，对牛和兔的心房MHC具有特异性。酶联免疫吸附竞争试验表明，这三种单克隆抗体识别抗原分子上不同的表位，尽管FA1和FA2的表位之间存在部分重叠。这些抗MHC单克隆抗体在研究心肌发育过程中MHC的表达方面将非常有用。
